The goal of Project 2 is to continue to provide the laboratory and preclinical experimental support for the ongoing clinical trials and to develop new ideas for future clinical trial approaches for malignant mesothelioma. The overall hypotheses of this continuing project is that immuno-gene therapy using adoptive T cell transfer can be used to treat malignant mesothelioma successfully, however this will require optimization to augment safety and efficacy. In this renewal, the previous preclinical focus on adenovirus interferon (now successfully being tested in a Phase 2 clinical trial in Project 1) will be redirected toward optimizing adoptively transferred human T-cells that express chimeric antigen receptors (CARs) to support our new and future clinical trials. Adoptive T cell transfer has shown great potential but faces a number of obstacles. Experiments proposed in this Project will focus on increasing trafficking of T cells into tumors, optimizing combination with chemotherapy, and enhancing T cell function by altering the tumor microenvironment and preventing T cell inactivation. Project 3 will explore other issues. In Specific Aim 1, the approaches and mechanisms of combining blockade of tumor-induced immunosuppression with a cycloxygenase-2 inhibitor with adoptive transfer of CAR-expressing T-cells in mesothelioma will be explored. In Specific Aim 2, approaches and mechanisms of combining chemotherapy with adoptive transfer therapy will be studied in both in immunodeficient and immunocompetent animal models. In Specific Aim 3, ways to improve trafficking of CARs will be investigated by genetically altering the T-cells to express new chemokine receptors to enhance T-cell trafficking. In Specific Aim 4, CAR-expressing T cells will be altered genetically to resist inactivation within the tumor by inhibiting the function of key inhibitory enzymes with an initial focus on SHP-1. Completion of these aims will: 1) provide key preclinical translational studies that can be rapidly applied to our evolving mesothelioma clinical trials in Project 1, 2) advance the entire field of adoptive T cell transfer by overcoming key obstacles, and 3) generate new knowledge about tumor immunology (especially in vivo human T cell biology).